Background and purpose: Perturbation of the endoplasmic reticulum (ER) homeostasis is known as one of the prominent features of obesity. Combining a healthy diet and exercise can improve health and performance. The aim of this study was to investigate the effect of Pilates training (PT) and resveratrol (RSV) on endoplasmic reticulum stress indicators in middle-aged obese women. Materials and methods: In this clinical trial, 40 obese adult women with the mean age of 42. 74±5. 70 years and body mass index of 31. 76±1. 37 kg/m2 in Tehran were randomly allocated into four groups,Placebo (C), Pilates Training (PT), Resveratrol (RSV), and Pilates Training-Resveratrol (PTRSV). The PT groups did the training program with 14-16 RPE for 8 weeks, 3 times/week. The RSV and PTRSV groups were provided with 500 mg RSV tablets every morning. Data were analyzed using independent t-test and ANCOVA. Results: According to findings, PT and RSV decreased Glucose-regulating protein 78 (GRP78) (P=0. 001 and P=0. 007, respectively) and Tribbles homolog 3 (TRB3) (P=0. 0001 and P=0. 0001, respectively). Simultaneous intervention of PT and RSV also reduced GRP78 and TRB3 more than other groups (P<0. 05). Conclusion: PT and RSV supplementation could improve the endoplasmic reticulum stress in middle-aged obese women by altering the levels of GRP78 and TRB3. However, the simultaneous effect of PT and RSV was found to be better on these indices.

Background: Multiple Sclerosis (MS) with four different types is one of the well studied autoimmune diseases of the central nervous system. Generally, two-thirds of MS patients are females who are at risk of pregnancy-related complications. Inappropriate responses of mother’ s immune system, such as antibody production against placental proteins, may lead to pregnancy-related disorders. The association between pregnancy complications and some autoantibodies including anti-phospholipid and anti-angiotensin II type-1 receptor antibodies are clear examples in this regard. Objective: To investigate the probable placental antigens that might be targeted by the antibodies in the sera of MS patients. Methods: Total placental proteins were extracted from normal fresh placentas and were separated using two-dimensional gel electrophoresis (2-DE) technique. The separated proteins were transferred onto a Polyvinylidene Fluoride (PVDF) membrane and blotted with the pooled sera of MS women or healthy controls (20 individuals in each group). The differentially blotted spot was identified by mass spectrometry and confirmed by western blot technique. Results: The results indicated that the women afflicted with MS had an antibody against placental HSP70kDa protein 5 (GRP78). Conclusion: In the present study, a new placental autoantigen candidate, which was targeted by antibody present in MS women sera, was found. The clinical importance of this finding regarding pregnancy complications in MS patients should be investigated by further experiments.

Introduction: Recurrent pregnancy loss (RPL) is defined as three or more miscarriages prior to the 20th week of gestation. This condition occurs in 15-20% of all pregnancies. Glucose-regulated protein 78 (GRP78) is an endoplasmic reticulum (ER) protein that is expressed on the trophoblast cells. The purpose of this study was to evaluate the presence of anti-GRP78 antibodies in the sera of women with RPL. Materials & Methods: This case-control study was conducted 80 women referring to the Infertility Center of Shiraz University of Medical Sciences, Shiraz, Iran, during 2014-2016. The study population was assigned into two groups of case entailing 38 women with unexplained RPL and control including 42 healthy pregnant women with no history of miscarriage and a minimum of two successful pregnancies. The measurement of the anti-GRP78 antibody in the sera of the two groups was performed using the enzyme-linked immunosorbent assay (ELISA). Furthermore, western blot technique was used to assess the expression of GRP78 in placenta tissue for both groups. Findings: The results of the western blot demonstrated the expression of GRP78 on the trophoblast cells in both groups. Furthermore, anti-GRP78 antibodies were detected in both case and control groups through ELISA. The case group had a lower level of anti-GRP78 antibodies; however, this difference was not statistically significant (P=0. 1). Discussion & Conclusions: The findings of the current study revealed no significant relationship between anti-GRP78 antibody level in RPL patients and healthy pregnant subjects.

Objective(s): Exosomes are nanovesicular vehicles capable of transporting different cargoes. Based on their characteristics, exosomes are proposed as a class of vehicles for targeted delivery of therapeutics. We aimed to establish a HEK293T stable cell line capable of secreting GRP78-specific scFv-targeted exosomes.Methods: The pLEX-LAMP2b-GRP78 construct was developed by enzymatic replacement of DARPin in pLEX-LAMP2b-DARPin with a GRP78-specific scFv. pLEX-LAMP2b-GRP78 (or TurboGFP as control), psPAX2, and pMD2.G plasmids were co-transfected into HEK293T cells, and produced lentiviruses were harvested. Different multiplicities of infection (MOI; 10, 20, 30, 60, 120, and 240) were used for the transduction of HEK293T to select the most appropriate one as assessed by flow cytometry. Transduced HEK293T cells were subject to puromycin selection and the presence of the scFv was assessed in the established cell line at the DNA, transcript, and protein levels by PCR, RT-PCR, and Western blotting, respectively.Results: pLEX-LAMP2b-GRP78 was successfully developed. Co-transfection resulted in the expression of GFP by HEK293T in the control group 48 hours following transfection. The MOI of 60 was selected as 10% of cells were GFP+ 72 hours following transduction. Following puromycin selection, the presence of the integrated scFv DNA and transcript was confirmed. Moreover, Western blotting results confirmed the presence of the His-tagged scFv in the established cell line.Conclusions: HEK293T cells can be engineered for the production of targeted exosomes which could be applied for therapeutic purposes. Moreover, scFvs are potent targeting domains that could be leveraged for the development of targeted exosomes.